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Fig. 6 | Molecular Cancer

Fig. 6

From: A novel mechanism of lncRNA and miRNA interaction: CCAT2 regulates miR-145 expression by suppressing its maturation process in colon cancer cells

Fig. 6

CCAT2 regulates miR-145 expression by directly suppressing its processing. Digoxigenin labeled 200 ng of pri-miR-145 (668 base) containing 1 μg of total RNAs from the CCAT2 overexpressing or knockout (KO) colon cancer cells was incubated at 25 °C for 5 min; 1unit of recombinant Dicer was added and incubated at 37 °C for 60 min. The samples were divided into two parts: (a) performed RT-PCR to detect CCAT2, pri-, pre- and mature miR-145. Agarose (3%) chromatography demonstrates changes in PCR products. (b) carried out electrophoresis on 8% polyacrylamide-8M urea gel, subsequently transferred onto membrane and the Dig-labeled RNAs detected with anti-digoxigenin antibody. M: 100 bp DNA (A) or RNA ladder (B), Lane-1: pri-miR-145, Lane-2: pri-miR-145 + Dicer, Lane-3: pri-miR-145 + RNA of CR-HT29 + Dicer, Lane-4: pri-miR-145 + RNA of CR-HT29/CCAT2 + Dicer, Lane-5: pri-miR-145 + RNA of HCT-116 + Dicer, Lane-6: pri-miR-145 + RNA of HCT-116/CCAT2 + Dicer, Lane-7: pri-miR-145 + RNA of HCT-116/KO-CCAT2 + Dicer

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